Suganuma Masatoshi, Nomura Tsuyoshi, Higa Yukiko, Kataoka Yukiko, Funaguma Shunsuke, Okazaki Hironobu, Suzuki Takeo, Fujiyama Kazuhito, Sezutsu Hideki, Tatematsu Ken-Ichiro, Tamura Toshiki
Sysmex Corporation, 4-4-4 Takatsukadai, Nishi-ku, Kobe, Hyogo 651-2271, Japan.
Sysmex Corporation, 4-4-4 Takatsukadai, Nishi-ku, Kobe, Hyogo 651-2271, Japan.
J Biosci Bioeng. 2018 Jul;126(1):9-14. doi: 10.1016/j.jbiosc.2018.01.007. Epub 2018 Feb 9.
A silkworm-baculovirus system is particularly effective for producing recombinant proteins, including glycoproteins. However, N-glycan structures in silkworm differ from those in mammals. Glycoproteins in silkworm are secreted as pauci-mannose type N-glycans without sialic acid or galactose residues. Sialic acid on N-glycans plays important roles in protein functions. Therefore, we developed pathways for galactosylation and sialylation in silkworm. Sialylated N-glycans on proteins were successfully produced in silkworm by co-expressing galactosyltransferase and sialyltransferase and providing an external supply of a sialylation-related substrate. α2,3/α2,6 Sialylation to N-glycans was controlled by changing the type of sialyltransferase expressed in silkworm. Furthermore, the co-expression of N-acetylglucosaminyltransferase II facilitated the formation of additional di-sialylated N-glycan structures. Our results provide new information on the control of N-glycosylation in silkworm.
家蚕杆状病毒系统在生产包括糖蛋白在内的重组蛋白方面特别有效。然而，家蚕中的N-聚糖结构与哺乳动物中的不同。家蚕中的糖蛋白以缺乏唾液酸或半乳糖残基的寡甘露糖型N-聚糖形式分泌。N-聚糖上的唾液酸在蛋白质功能中起重要作用。因此，我们在家蚕中开发了半乳糖基化和唾液酸化途径。通过共表达半乳糖基转移酶和唾液酸转移酶并提供唾液酸化相关底物的外部供应，在家蚕中成功产生了蛋白质上的唾液酸化N-聚糖。通过改变家蚕中表达的唾液酸转移酶的类型来控制N-聚糖的α2,3/α2,6唾液酸化。此外，N-乙酰葡糖胺转移酶II的共表达促进了额外的二唾液酸化N-聚糖结构的形成。我们的结果提供了有关家蚕中N-糖基化控制的新信息。
